LOCUS       sequence-392399-       11573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for genomic targeting of a Tetracycline-inducible Cas9 
            cassette to the human AAVS1/PPP1R12C locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11573)
  AUTHORS   Gonzalez F, Zhu Z, Shi ZD, Lelli K, Verma N, Li QV, Huangfu D
  TITLE     An iCRISPR Platform for Rapid, Multiplexable, and Inducible Genome 
            Editing in Human Pluripotent Stem Cells.
  JOURNAL   Cell Stem Cell. 2014 Jun 11. pii: S1934-5909(14)00205-7. doi: 
            10.1016/j.stem.2014.05.018.
  PUBMED    24931489
REFERENCE   2  (bases 1 to 11573)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Stem 
            Cell. 2014 Jun 11. pii: S1934-5909(14)00205-7. doi: 
            10.1016/j.stem.2014.05.018."
COMMENT     Sequence Label: Puro-Cas9 donor
FEATURES             Location/Qualifiers
     source          1..11573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      40..628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     529..548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     782..799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    916..937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(952..957,958..975,976..982)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    990..1006
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     995..1017
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1014..1030
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1014..1030
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1051..1069
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    1104..1907
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    1914..1939
                     /label=SA
                     /note="splice acceptor site"
     CDS             1963..2016
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2026..2625
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2026..2045)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2522..2542
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2657..2674)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2663..2887
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2894..2975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2919..2938
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    3159..3183
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3209..3256)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(3257..7357)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(3257..7357)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(7382..7402)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7409..7474)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      7474..7483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(7508..7532)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(7549..7863)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(7552..7576)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    7623..7641
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7659..7677
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7694..7712
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7730..7748
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7766..7784
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7801..7819
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7837..7855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    7880..8716
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(8750..8769)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8751..8769)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8776..8793)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8776..8792)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8785..8807)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8930..9232
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     9111..9130
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(9635..9654)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     10249..10268
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(10629..11420,11421..11489))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     11252..11271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
       61 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      181 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      781 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
      841 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      901 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
      961 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     1021 cagctatgac catgattacg ccaagctcag aattaaccct cactaaaggg actagtcctg
     1081 caggtttaaa cgaattcgcc ctttgctttc tctgaccagc attctctccc ctgggcctgt
     1141 gccgctttct gtctgcagct tgtggcctgg gtcacctcta cggctggccc agatccttcc
     1201 ctgccgcctc cttcaggttc cgtcttcctc cactccctct tccccttgct ctctgctgtg
     1261 ttgctgccca aggatgctct ttccggagca cttccttctc ggcgctgcac cacgtgatgt
     1321 cctctgagcg gatcctcccc gtgtctgggt cctctccggg catctctcct ccctcaccca
     1381 accccatgcc gtcttcactc gctgggttcc cttttccttc tccttctggg gcctgtgcca
     1441 tctctcgttt cttaggatgg ccttctccga cggatgtctc ccttgcgtcc cgcctcccct
     1501 tcttgtaggc ctgcatcatc accgtttttc tggacaaccc caaagtaccc cgtctccctg
     1561 gctttagcca cctctccatc ctcttgcttt ctttgcctgg acaccccgtt ctcctgtgga
     1621 ttcgggtcac ctctcactcc tttcatttgg gcagctcccc tacccccctt acctctctag
     1681 tctgtgctag ctcttccagc cccctgtcat ggcatcttcc aggggtccga gagctcagct
     1741 agtcttcttc ctccaacccg ggcccctatg tccacttcag gacagcatgt ttgctgcctc
     1801 cagggatcct gtgtccccga gctgggacca ccttatattc ccagggccgg ttaatgtggc
     1861 tctggttctg ggtactttta tctgtcccct ccaccccaca gtggggcaag cttctgacct
     1921 cttctcttcc tcccacaggg cctcgagaga tctggcagcg gagagggcag aggaagtctt
     1981 ctaacatgcg gtgacgtgga ggagaatccc ggccctaggc tcgagatgac cgagtacaag
     2041 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     2101 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     2161 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     2221 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     2281 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     2341 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     2401 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     2461 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     2521 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     2581 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgatctag agggcccgtt
     2641 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2701 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2761 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2821 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2881 tctatgggtc gaggcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2941 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     3001 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     3061 gtggtatggc tgattatgat cctctagaga tatcgtcgac aagcttatcg atgcggccgc
     3121 gctagcacgc gtcagctgac tagaggatcc ccggatcaac cactttgtac aagaaagctg
     3181 ggtcggcgcg cccacccttg aattcttact ttttcttttt tgcctggccg gcctttttcg
     3241 tggccgccgg ccttttgtcg cctcccagct gagacaggtc gatccgtgtc tcgtacaggc
     3301 cggtgatgct ctggtggatc agggtggcgt ccagcacctc tttggtgctg gtgtacctct
     3361 tccggtcgat ggtggtgtca aagtacttga aggcggcagg ggctcccaga ttggtcaggg
     3421 taaacaggtg gatgatattc tcggcctgct ctctgatggg cttatcccgg tgcttgttgt
     3481 aggcggacag cactttgtcc agattagcgt cggccaggat cactctcttg gagaactcgc
     3541 tgatctgctc gatgatctcg tccaggtagt gcttgtgctg ttccacaaac agctgtttct
     3601 gctcattatc ctcgggggag cccttcagct tctcatagtg gctggccagg tacaggaagt
     3661 tcacatattt ggagggcagg gccagttcgt ttcccttctg cagttcgccg gcagaggcca
     3721 gcattctctt ccggccgttt tccagctcga acagggagta cttaggcagc ttgatgatca
     3781 ggtccttttt cacttctttg tagcccttgg cttccagaaa gtcgatggga ttcttctcga
     3841 agctgcttct ttccatgatg gtgatcccca gcagctcttt cacactcttc agtttcttgg
     3901 acttgccctt ttccactttg gccaccacca gcacagaata ggccacggtg gggctgtcga
     3961 agccgccgta cttcttaggg tcccagtcct tctttctggc gatcagctta tcgctgttcc
     4021 tcttgggcag gatagactct ttgctgaagc cgcctgtctg cacctcggtc tttttcacga
     4081 tattcacttg gggcatgctc agcactttcc gcacggtggc aaaatcccgg cccttatccc
     4141 acacgatctc cccggtttcg ccgtttgtct cgatcagagg ccgcttccgg atctcgccgt
     4201 tggccagggt aatctcggtc ttgaaaaagt tcatgatgtt gctgtagaag aagtacttgg
     4261 cggtagcctt gccgatttcc tgctcgctct tggcgatcat cttccgcacg tcgtacacct
     4321 tgtagtcgcc gtacacgaac tcgctttcca gcttagggta ctttttgatc agggcggttc
     4381 ccacgacggc gttcaggtag gcgtcgtggg cgtggtggta gttgttgatc tcgcgcactt
     4441 tgtaaaactg gaaatccttc cggaaatcgg acaccagctt ggacttcagg gtgatcactt
     4501 tcacttcccg gatcagcttg tcattctcgt cgtacttagt gttcatccgg gagtccagga
     4561 tctgtgccac gtgctttgtg atctgccggg tttccaccag ctgtctcttg atgaagccgg
     4621 ccttatccag ttcgctcagg ccgcctctct cggccttggt cagattgtcg aactttctct
     4681 gggtaatcag cttggcgttc agcagctgcc gccagtagtt cttcatcttc ttcacgacct
     4741 cttcggaggg cacgttgtcg ctcttgcccc ggttcttgtc gcttctggtc agcaccttgt
     4801 tgtcgatgga gtcgtccttc agaaagctct gaggcacgat atggtccaca tcgtagtcgg
     4861 acagccggtt gatgtccagt tcctggtcca cgtacatatc ccgcccattc tgcaggtagt
     4921 acaggtacag cttctcgttc tgcagctggg tgttttccac ggggtgttct ttcaggatct
     4981 ggctgcccag ctctttgatg ccctcttcga tccgcttcat tctctcgcgg ctgttcttct
     5041 gtcccttctg ggtggtctgg ttctctctgg ccatttcgat cacgatgttc tcgggcttgt
     5101 gccggcccat cactttcacg agctcgtcca ccaccttcac tgtctgcagg atgcccttct
     5161 taatggcggg gctgccggcc agattggcaa tgtgctcgtg caggctatcg ccctggccgg
     5221 acacctgggc tttctggatg tcctctttaa aggtcaggct gtcgtcgtgg atcagctgca
     5281 tgaagtttct gttggcgaag ccgtcggact tcaggaaatc caggattgtc ttgccggact
     5341 gcttgtcccg gatgccgttg atcagcttcc ggctcagcct gccccagccg gtgtatctcc
     5401 gccgcttcag ctgcttcatc actttgtcgt cgaacaggtg ggcataggtt ttcagccgtt
     5461 cctcgatcat ctctctgtcc tcaaacagtg tcagggtcag cacgatatct tccagaatgt
     5521 cctcgttttc ctcattgtcc aggaagtcct tgtccttgat aattttcagc agatcgtggt
     5581 atgtgcccag ggaggcgttg aaccgatctt ccacgccgga gatttccacg gagtcgaagc
     5641 actcgatttt cttgaagtag tcctctttca gctgcttcac ggtcactttc cggttggtct
     5701 tgaacagcag gtccacgatg gcctttttct gctcgccgct caggaaggcg ggctttctca
     5761 ttccctcggt cacgtatttc actttggtca gctcgttata cacggtgaag tactcgtaca
     5821 gcaggctgtg cttgggcagc accttctcgt tgggcaggtt cttatcgaag ttggtcatcc
     5881 gctcgatgaa gctctgggcg gaagcgccct tgtccaccac ttcctcgaag ttccaggggg
     5941 tgatggtttc ctcgctcttt ctggtcatcc aggcgaatct gctgtttccc ctggccagag
     6001 ggcccacgta gtaggggatg cggaaggtca ggatcttctc gatcttttcc cggttgtcct
     6061 tcaggaatgg gtaaaaatct tcctgccgcc gcagaatggc gtgcagctct cccaggtgga
     6121 tctggtgggg gatgctgccg ttgtcgaagg tccgctgctt ccgcagcagg tcctctctgt
     6181 tcagcttcac gagcagttcc tcggtgccgt ccatcttttc caggatgggc ttgatgaact
     6241 tgtagaactc ttcctggctg gctccgccgt caatgtagcc ggcgtagccg ttcttgctct
     6301 ggtcgaagaa aatctctttg tacttctcag gcagctgctg ccgcacgaga gctttcagca
     6361 gggtcaggtc ctggtggtgc tcgtcgtatc tcttgatcat agaggcgctc aggggggcct
     6421 tggtgatctc ggtgttcact ctcaggatgt cgctcagcag gatggcgtcg gacaggttct
     6481 tggcggccag aaacaggtcg gcgtactggt cgccgatctg ggccagcagg ttgtccaggt
     6541 cgtcgtcgta ggtgtccttg ctcagctgca gtttggcatc ctcggccagg tcgaagttgc
     6601 tcttgaagtt gggggtcagg cccaggctca gggcaatcag gtttccgaac aggccattct
     6661 tcttctcgcc gggcagctgg gcgatcagat tttccagccg tctgctcttg ctcagtctgg
     6721 cagacaggat ggccttggcg tccacgccgc tggcgttgat ggggttttcc tcgaacagct
     6781 ggttgtaggt ctgcaccagc tggatgaaca gcttgtccac gtcgctgttg tcggggttca
     6841 ggtcgccctc gatcaggaag tggccccgga acttgatcat gtgggccagg gccagataga
     6901 tcagccgcag gtcggccttg tcggtgctgt ccaccagttt ctttctcagg tggtagatgg
     6961 tggggtactt ctcgtggtag gccacctcgt ccacgatgtt gccgaagatg gggtgccgct
     7021 cgtgcttctt atcctcttcc accaggaagg actcttccag tctgtggaag aagctgtcgt
     7081 ccaccttggc catctcgttg ctgaagatct cttgcagata gcagatccgg ttcttccgtc
     7141 tggtgtatct tcttctggcg gttctcttca gccgggtggc ctcggctgtt tcgccgctgt
     7201 cgaacagcag ggctccgatc aggttcttct tgatgctgtg ccggtcggtg ttgcccagca
     7261 ccttgaattt cttgctgggc accttgtact cgtcggtgat cacggcccag cccacagagt
     7321 tggtgccgat gtccaggccg atgctgtact tcttgtcggc tgctgggact ccgtggatac
     7381 cgaccttccg cttcttcttt ggggccatct tatcgtcatc gtctttgtaa tcaatatcat
     7441 gatccttgta gtctccgtcg tggtccttat agtccatggt ggcaccggtc caacctgaaa
     7501 accgcggagc ctgctttttt gtacaaactt gttgatcgga attctccagg cgatctgacg
     7561 gttcactaaa cgagctctgc ttatataggc ctcccaccgt acacgcctac ctcgacatac
     7621 gttctctatc actgataggg agtaaactcg acatacgttc tctatcactg atagggataa
     7681 actcgacata cgttctctat cactgatagg gagtaaactc gacatacgtt ctctatcact
     7741 gatagggagt aaactcgaca tacgttctct atcactgata gggagtaaac tcgacatcgt
     7801 tctctatcac tgatagggag taaactcgac atacgttctc tatcactgat agggagtaaa
     7861 ctcgacagta ctaagcttta ctagggacag gattggtgac agaaaagccc catccttagg
     7921 cctcctcctt cctagtctcc tgatattggg tctaaccccc acctcctgtt aggcagattc
     7981 cttatctggt gacacacccc catttcctgg agccatctct ctccttgcca gaacctctaa
     8041 ggtttgctta cgatggagcc agagaggatc ctgggaggga gagcttggca gggggtggga
     8101 gggaaggggg ggatgcgtga cctgcccggt tctcagtggc caccctgcgc taccctctcc
     8161 cagaacctga gctgctctga cgcggctgtc tggtgcgttt cactgatcct ggtgctgcag
     8221 cttccttaca cttcccaaga ggagaagcag tttggaaaaa caaaatcaga ataagttggt
     8281 cctgagttct aactttggct cttcaccttt ctagtcccca atttatattg ttcctccgtg
     8341 cgtcagtttt acctgtgaga taaggccagt agccagcccc gtcctggcag ggctgtggtg
     8401 aggagggggg tgtccgtgtg gaaaactccc tttgtgagaa tggtgcgtcc taggtgttca
     8461 ccaggtcgtg gccgcctcta ctccctttct ctttctccat ccttctttcc ttaaagagtc
     8521 cccagtgcta tctgggacat attcctccgc ccagagcagg gtcccgcttc cctaaggccc
     8581 tgctctgggc ttctgggttt gagtccttgg caagcccagg agaggcgctc aggcttccct
     8641 gtcccccttc ctcgtccacc atctcatgcc cctggctctc ctgccccttc cctacagggg
     8701 ttcctggctc tgctctaagg gcgaattcgc ggccgctaaa ttcaattcgc cctatagtga
     8761 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     8821 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     8881 ggcccgcacc gatcgccctt cccaacagtt gcgcagccta tacgtacggc agtttaaggt
     8941 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     9001 tgacacgccg gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     9061 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     9121 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     9181 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     9241 catgagatta tcaaaaagga tcttcaccta gatccttttc acgtagaaag ccagtccgca
     9301 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
     9361 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
     9421 ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
     9481 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     9541 atcaagctct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     9601 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     9661 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     9721 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     9781 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     9841 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     9901 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     9961 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
    10021 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
    10081 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
    10141 ccatgcatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
    10201 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
    10261 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
    10321 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa
    10381 ttattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
    10441 cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
    10501 ttctaaatac attcaaatat gtatccgctc atgagattat caaaaaggat cttcacctag
    10561 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
    10621 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
    10681 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
    10741 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
    10801 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
    10861 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
    10921 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
    10981 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
    11041 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
    11101 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
    11161 tgcttttctg tgactggtga atactcaacc aagtcattct gagaatagtg tatgcggcga
    11221 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
    11281 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    11341 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    11401 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    11461 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    11521 tatcagggtt attgtctcat gaccaaaatc ccttaacgtg agttttcgtt cca
//
